Molecular genetic study on the metabolism and function of phosphatidylserine in mammalian cells

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To reveal the mechanism of phosphatidylserine (PS) biosynthesis and function of PS, we previously isolated a mammalian cell mutant defective in PS biosyntheses. By using this mutant, we found that PS synthase and PS decarboxylase played important roles in PS formation in mammalian cells. In the present study, to further characterize the molecular mechanism of PS synthesis, we tried to purify PS synthase and to isolate mutants defective in PS decarboxylase.(1) We made polyclonal antibody to PS synthase by using a oligopeptide which corresponded to the C-terminal part of PS synthase.(2) We tried to solubilize PS synthase by using various kinds of detergents. However, we could not find good conditions to solubilize the enzyme. (3) We desrupted the gene of PS decarboxylase in CHO cells by gene targeting. In this way, we obtained a mutant defective in one of the Ps decarboxylase genes.Our results are still premature and so we will continue the above studies for coming several years.